Synthesis of spin-labeled riboswitch RNAs using convertible nucleosides and DNA-catalyzed RNA ligation
Synthesis of spin-labeled riboswitch RNAs using convertible nucleosides and DNA-catalyzed RNA ligation
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DOI：
10.1016/j.bmc.2013.04.007
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发表时间：
2013-10-15
影响因子：
3.5
通讯作者：
Hoebartner, Claudia
中科院分区：
文献类型：
--
作者：
Buettner, Lea;Seikowski, Jan;Hoebartner, Claudia
可以通过电子顺磁共振 (EPR) 光谱监测的化学稳定的硝基氧自由基可以提供有关核糖开关等功能性 RNA 的结构和动态特性的信息。可转换核苷方法用于在 RNA 中胞苷和 2'-O-甲基胞苷核苷酸的环外 N-4-氨基上安装 2,2,6,6-四甲基哌啶-1-氧基 (TEMPO) 和 2,2,5,5-四甲基吡咯烷-1-氧基（丙氧基）标记。为了获得超出固相合成极限的位点特异性标记的长核糖开关RNA，我们报告了使用体外选择的脱氧核酶作为催化剂连接自旋标记的RNA，并证明了TEMPO标记的53 nt SAM-III和118 nt SAM-I核糖开关结构域（SAM = S-腺苷甲硫氨酸）的合成。 (C) 2013 Elsevier Ltd. 保留所有权利。
Chemically stable nitroxide radicals that can be monitored by electron paramagnetic resonance (EPR) spectroscopy can provide information on structural and dynamic properties of functional RNA such as riboswitches. The convertible nucleoside approach is used to install 2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPO) and 2,2,5,5-tetramethylpyrrolidin-1-oxyl (proxyl) labels at the exocyclic N-4-amino group of cytidine and 2'-O-methylcytidine nucleotides in RNA. To obtain site-specifically labeled long riboswitch RNAs beyond the limit of solid-phase synthesis, we report the ligation of spin-labeled RNA using an in vitro selected deoxyribozyme as catalyst, and demonstrate the synthesis of TEMPO-labeled 53 nt SAM-III and 118 nt SAM-I riboswitch domains (SAM = S-adenosylmethionine). (C) 2013 Elsevier Ltd. All rights reserved.